Cytochrome P450 Function Research Articles

Identification of Electron Transfer in the System of Ferredoxins and Ferredoxin Reductases from Mycolicibacterium smegmatis

Steroid-26-monooxygenases belong to the cytochrome P450 superfamily and function as part of three-component systems together with ferredoxins and ferredoxin reductases providing electron transport. The P450-dependent redox partners of the actinobacterial strain Mycolicibacterium smegmatis mc2155 were investigated. The genes encoding mycolibacterial ferredoxins (FdxD and FdxE) and ferredoxin reductases (FdrA and FprA) were overexpressed in E. coli cells. A scheme for isolation and purification of synthesized recombinant proteins using affinity chromatography was developed, resulting in electrophoretically homogeneous preparations. Spectral analysis of ferredoxin reductase showed absorption peaks characteristic of FAD-containing proteins. The reaction of cytochrome c reduction using recombinant proteins was carried out, demonstrating that FdxD, FdxE, FdrA, and FprA can act as components of electron transport from the reducing equivalents of NAD(P)H.

Harnessing Porphyrin Accumulation in Liver Cancer: Combining Genomic Data and Drug Targeting.

The liver, a pivotal organ in human metabolism, serves as a primary site for heme biosynthesis, alongside bone marrow. Maintaining precise control over heme production is paramount in healthy livers to meet high metabolic demands while averting potential toxicity from intermediate metabolites, notably protoporphyrin IX. Intriguingly, our recent research uncovers a disrupted heme biosynthesis process termed 'porphyrin overdrive' in cancers that fosters the accumulation of heme intermediates, potentially bolstering tumor survival. Here, we investigate heme and porphyrin metabolism in both healthy and oncogenic human livers, utilizing primary human liver transcriptomics and single-cell RNA sequencing (scRNAseq). Our investigations unveil robust gene expression patterns in heme biosynthesis in healthy livers, supporting electron transport chain (ETC) and cytochrome P450 function without intermediate accumulation. Conversely, liver cancers exhibit rewired heme biosynthesis and a massive downregulation of cytochrome P450 gene expression. Notably, despite diminished drug metabolism, gene expression analysis shows that heme supply to the ETC remains largely unaltered or even elevated with patient cancer progression, suggesting a metabolic priority shift. Liver cancers selectively accumulate intermediates, which are absent in normal tissues, implicating their role in disease advancement as inferred by expression analysis. Furthermore, our findings in genomics establish a link between the aberrant gene expression of porphyrin metabolism and inferior overall survival in aggressive cancers, indicating potential targets for clinical therapy development. We provide in vitro proof-of-concept data on targeting porphyrin overdrive with a drug synergy strategy.

A physiologically-based pharmacokinetic modeling approach for dosing amiodarone in children on ECMO.

Extracorporeal membrane oxygenation (ECMO) is a cardiopulmonary bypass device commonly used to treat cardiac arrest in children. The American Heart Association guidelines for cardiopulmonary resuscitation (CPR) and emergency cardiovascular care recommend using amiodarone as a first-line agent to treat ventricular arrhythmias in children with cardiac arrest. However, there are no dosing recommendations for amiodarone to treat ventricular arrhythmias in pediatric patients on ECMO. Amiodarone has a high propensity for adsorption to the ECMO components due to its physicochemical properties leading to altered pharmacokinetics (PK) in ECMO patients. The change in amiodarone PK due to interaction with ECMO components may result in a difference in optimal dosing in patients on ECMO when compared with non-ECMO patients. To address this clinical knowledge gap, a physiologically-based pharmacokinetic model of amiodarone was developed in adults and scaled to children, followed by the addition of an ECMO compartment. The pediatric model included ontogeny functions of cytochrome P450 (CYP450) enzyme maturation across various age groups. The ECMO compartment was parameterized using the adsorption data of amiodarone obtained from exvivo studies. Model predictions captured observed concentrations of amiodarone in pediatric patients with ECMO well with an average fold error between 0.5 and 2. Model simulations support an amiodarone intravenous (i.v) bolus dose of 22 mg/kg (neonates), 13 mg/kg (infants), 8 mg/kg (children), and 6 mg/kg (adolescents). This PBPK modeling approach can be applied to explore the dosing of other drugs used in children on ECMO.

Hordeum vulgareCYP76M57 catalyzes C2 shortening of tryptophan side chain by C-N bond rearrangement in gramine biosynthesis.

The indole alkaloid gramine, 3-(dimethylaminomethyl)indole, is a defensive specialized metabolite found in some barley cultivars. In its biosynthetic process, the tryptophan (Trp) side chain is shortened by two carbon atoms to produce 3-(aminomethyl)indole (AMI), which is then methylated by N-methyltransferase (HvNMT) to produce gramine. Although side chain shortening is one of the crucial scaffold formation steps of alkaloids originating from aromatic amino acids, the gene and enzyme involved in the Trp-AMI conversion reactions are unknown. In this study, through RNA-seq analysis, 35 transcripts were shown to correlate with gramine production; among them, an uncharacterized cytochrome P450 (CYP) gene, CYP76M57, and HvNMT were identified as candidate genes for gramine production. Transgenic Arabidopsis thaliana and rice overexpressing CYP and HvNMT accumulate AMI, N-methyl-AMI, and gramine. CYP76M57, heterologously expressed in Pichia pastoris, was able to act on Trp to produce AMI. Furthermore, the amino group nitrogen of Trp was retained during the CYP76M57-catalyzed reaction, indicating that the C2 shortening of Trp proceeds with an unprecedented biosynthetic process, the removal of the carboxyl group and Cα and the rearrangement of the nitrogen atom to Cβ. In some gramine-non-accumulating barley cultivars, arginine 104 in CYP76M57 is replaced by threonine, which abolished the catalytic activity of CYP76M57 to convert Trp into AMI. These results uncovered the missing committed enzyme of gramine biosynthesis in barley and contribute to the elucidation of the potential functions of CYPs in plants and undiscovered specialized pathways.

Genome-wide identification and expression analysis of cytochrome P450 gene family in Macrobrachium nipponense

Cytochrome P450 (CYP450) is a superfamily widely found in various biological organisms, including animals, plants, and microorganisms. CYP450 is involved in the metabolism of endogenous and exogenous substances. In addition, a few CYP450 genes named as Halloween genes (CYP302a1, CYP307a1, etc.) are associated with the synthesis and secretion of ecdysone in Arthropods. Macrobrachium nipponense (Arthropoda, Crustacean) has many excellent breeding characteristics; for example, rapid growth rate, ability to survive in a variety of environments and high economic value. In order to further explored the structure and function of CYP450, a genome-wide analysis of CYP450 gene family was performed in M. nipponense. A total of 67 CYP450 genes were identified, and extensive bioinformatic analysis methods were conducted, including physicochemical property, sequence alignment, evolutionary tree construction, conserved sequence, chromosome localization, protein-protein interactions, and quantitative expression. The results showed that most CYP450 genes being around 500 amino acids, with 25 tandem repeats genes. The majority of CYP450 genes have the signature structure of the CYP450 gene family to ensure the stability of the spatial structure and the ability to bind substrates. The Halloween genes and CYP18a1 sequences are similar and occupy an importance role in M. nipponense. Finally, most CYP450 genes were up-regulated in gill and hepatopancreas of M. nipponense under acute high temperature stress, with significant up-regulation of Halloween gene expression. This study could further deepen the understanding of the structure and function of CYP450 in M. nipponense and investigate the role of CYP450 in freshwater crustaceans under high temperatures.

Cytochrome P450 2B6 6 distribution among patients living with HIV in Burkina Faso

The function of cytochrome P450 (CYP) enzymes, one of the major catalysts in drug metabolism, is significantly influenced by genetic polymorphisms leading to substantial inter-individual variability in drug response and/ or adverse reactions activity. The aim of this study was to determine the frequency of the CYP2B6 516G>T allele in patients living with HIV receiving treatment at the Medical Center of Camp General Aboubacar Sangoule Lamizana for whom the treatment was changed after the initiation of treatment because of the appearance of side effects. 155 HIV-positive patients were enrolled. Genomic Deoxyribonucleic acid (DNA) extraction from blood was performed using the GenJET® Genomic DNA according to the manufacturer’s instructions. Genotyping for the cytochrome P450 variant alleles was performed using predesigned primers. The amplification was carried out by Polymerase Chain Reaction (PCR), while the differentiation between the alleles was carried out after digestion with restriction enzymes by electrophoresis. The results obtained showed. Among these patients, 61 changed treatments during their follow-up. The majority of the 61 patients, 32 patients, received an initial treatment regimen based on the AZT (3'-Azido-3'-Deoxythymidine)/3TC (Lamivudine)/ NVP(Nevirapine) combination, followed by 18 patients for the TDF (tenofovir)/3TC (lamivudine)/ EFV(Efavirenz) regimens. Currently, the majority of patients, 48 patients, have benefited from a therapeutic regimen based on the TDF (tenofovir)/3TC (lamivudine) /DTG(Dolutegravir) combination, followed by 11 patients for the TDF (tenofovir)/3TC (lamivudine)/EFV(Efavirenz) regimens. The distribution of the major allele CYP2B6*T was 68%, and the minor CYP2B6*G allele was 32%. Heterozygote (GT genotype) and Homozygous mutants (TT genotype) and Heterozygote were 31% and 52,5%. Studies could be continued to set up cytochrome mutation detection kits to anticipate the onset of side effects and adapt treatments.

The effect of brain serotonin deficit (TPH2-KO) on the expression and activity of liver cytochrome P450 enzymes in aging male Dark Agouti rats

BackgroundLiver cytochrome P450 (CYP) greatly contributes to the metabolism of endogenous substances and drugs. Recent studies have demonstrated that CYP expression in the liver is controlled by the central nervous system via hormonal pathways. In particular, the expression of hepatic CYPs is negatively regulated by the brain serotoninergic system. The present study aimed to investigate changes in the function of the main liver drug-metabolizing CYP enzymes as a result of serotonin depletion in the brain of aging rats, caused by knockout of brain tryptophan hydroxylase gene (TPH2-KO).MethodsThe hepatic CYP mRNA (qRT-PCR), protein level (Western blotting) and activity (HPLC), and serum hormone levels (ELISA) were measured in Dark Agouti wild-type (WT) male rats (mature 3.5-month-old and senescent 21-month-old) and in TPH2-KO senescent animals.ResultsThe expression/activity of the studied CYPs decreased with age in the liver of wild-type rats. The deprivation of serotonin in the brain of aging males decreased the mRNA level of most of the studied CYPs (CYP1A/2A/2B/3A), and lowered the protein level of CYP2C11 and CYP3A. In contrast, the activities of CYP2C11, CYP3A and CYP2C6 were increased. The expression of cytochrome b5 decreased in aging rats, but increased in TPH2-deficient senescent animals. The serum concentration of growth hormone declined in the aged and further dropped down in TPH2-deficient senescent rats.ConclusionsRat liver cytochrome P450 functions deteriorate with age, which may impair drug metabolism. The TPH2 knockout, which deprives brain serotonin, affects cytochrome P450 expression and activity differently in mature and senescent male rats.

Human Cytochrome P450 1, 2, 3 Families as Pharmacogenes with Emphases on Their Antimalarial and Antituberculosis Drugs and Prevalent African Alleles.

Precision medicine gives individuals tailored medical treatment, with the genotype determining the therapeutic strategy, the appropriate dosage, and the likelihood of benefit or toxicity. Cytochrome P450 (CYP) enzyme families 1, 2, and 3 play a pivotal role in eliminating most drugs. Factors that affect CYP function and expression have a major impact on treatment outcomes. Therefore, polymorphisms of these enzymes result in alleles with diverse enzymatic activity and drug metabolism phenotypes. Africa has the highest CYP genetic diversity and also the highest burden of malaria and tuberculosis, and this review presents current general information on CYP enzymes together with variation data concerning antimalarial and antituberculosis drugs, while focusing on the first three CYP families. Afrocentric alleles such as CYP2A6*17, CYP2A6*23, CYP2A6*25, CYP2A6*28, CYP2B6*6, CYP2B6*18, CYP2C8*2, CYP2C9*5, CYP2C9*8, CYP2C9*9, CYP2C19*9, CYP2C19*13, CYP2C19*15, CYP2D6*2, CYP2D6*17, CYP2D6*29, and CYP3A4*15 are implicated in diverse metabolic phenotypes of different antimalarials such as artesunate, mefloquine, quinine, primaquine, and chloroquine. Moreover, CYP3A4, CYP1A1, CYP2C8, CYP2C18, CYP2C19, CYP2J2, and CYP1B1 are implicated in the metabolism of some second-line antituberculosis drugs such as bedaquiline and linezolid. Drug-drug interactions, induction/inhibition, and enzyme polymorphisms that influence the metabolism of antituberculosis, antimalarial, and other drugs, are explored. Moreover, a mapping of Afrocentric missense mutations to CYP structures and a documentation of their known effects provided structural insights, as understanding the mechanism of action of these enzymes and how the different alleles influence enzyme function is invaluable to the advancement of precision medicine.

Cocaine-Induced Time-Dependent Alterations in Cytochrome P450 and Liver Function

Cytochrome P450 is responsible for the metabolism of endogenous substrates, drugs and substances of abuse. The brain and nervous system regulate liver cytochrome P450 via neuroendocrine mechanisms, as shown in rodents. Cocaine exerts its addictive effects through the dopaminergic system, the functioning of which undergoes changes during its continuous use. Therefore, it can be hypothesized that the regulation of cytochrome P450 by cocaine may also alter during the addiction process, cessation and relapse. We analyzed preclinical studies on the mechanisms of the pharmacological action of cocaine, the role of the brain's dopaminergic system in the neuroendocrine regulation of cytochrome P450 and the in vitro and in vivo effects of cocaine on the cytochrome P450 expression/activity and hepatotoxicity. The results of passive cocaine administration indicate that cocaine affects liver cytochrome P450 enzymes (including those engaged in its own metabolism) via different mechanisms involving the expression of genes encoding cytochrome P450 enzymes and interaction with enzyme proteins. Thus, it may affect its own oxidative metabolism and the metabolism of endogenous substrates and other co-administered drugs and may lead to hepatotoxicity. Its effect depends on the specific cytochrome P450 enzyme affected, cocaine dosage, treatment duration and animal species. However, further complementary studies are needed to find out whether cocaine affects cytochrome P450 via the brain's dopaminergic system. The knowledge of cocaine's effect on cytochrome P450 function during the entire addiction process is still incomplete. There is a lack of information on the enzyme expression/activity in animals self-administering cocaine (addicted), in those withdrawn after cocaine self-administration, and during relapse in animals previously addicted; furthermore, there is no such information concerning humans. The subject of cytochrome P450 regulation by cocaine during the addiction process is an open issue, and addressing this topic may help in the treatment of drug abuse patients.

A Beneficial Action of Ricinus Communis

The Ricinus communis is very valuable plant for the human beings. Ricinus communis also known as castor plant. It has high medicinal value as well as pharmacological value for disease cure activity as well as much more traditional value. In a Castor plant, oil seed with rich oil content shows its high phytochemical compound of monounsaturated fatty acid and bioactive compounds. The predominant triglyceride component in the oil is triricinolein. Minor biological compounds including carotenoid, tocopherol, tocotrienol, phytosterol, phospholipid, phytochemical, and phenolic compounds are present in castor oil. R. communis harbours phytochemicals which have been shown the many receptor activate like peroxisome proliferator activated receptor these are receptor are responsible for the transcription factors regulating the expression of genes., nuclear factor NF-k-B responsible for the a regulator of innate immunity, cytochrome P450 function is hemeprotein that plays a key role in the metabolism of drugs and other xenobiotics, P38 mitogen-activated protein kinases kinase (p38 MAPK), tumor protein P53, B-cell lymphoma-extra-large (Bcl-xL) and vascular endothelial growth factor receptor-2 (VEGFR-2)These compounds offer oxidation stability, anti-inflammatory, and antioxidant properties to the oil. Traditionally the plant is used as laxative, purgative, fertilizer and fungicide etc. whereas the plant possesses beneficial effects such as anti-oxidant, Antinociceptive, antiasthmatic, antiulcer, Antidiabetic, Antifertility etc medicinal properties. This activity show in plant possesses due to the Valuable phytochemical constituents like flavonoids, saponins, glycosides, alkaloids and steroids etc. The motive of this paper is to explain the details of phyto-pharmacological properties of R. communis for the future research work for the upcoming research scholar. Keywords: Ricinus communis, castor plant, antimicrobial and pharmacology.

Ebola virus infection induces a delayed type I IFN response in bystander cells and the shutdown of key liver genes in human iPSC-derived hepatocytes.

SummaryLiver damage and an exacerbated inflammatory response are hallmarks of Ebola virus (EBOV) infection. Little is known about the intrinsic response to infection in human hepatocytes and their contribution to inflammation. Here, we present an induced pluripotent stem cell (iPSC)-derived hepatocyte-like cell (HLC) platform to define the hepato-intrinsic response to EBOV infection. We used this platform to show robust EBOV infection, with characteristic ultrastructural changes and evidence for viral replication. Transcriptomics analysis revealed a delayed response with minimal early transcriptomic changes, followed by a general downregulation of hepatic function and upregulation of interferon signaling, providing a potential mechanism by which hepatocytes participate in disease severity and liver damage. Using RNA-fluorescence in situ hybridization (FISH), we showed that IFNB1 and CXCL10 were mainly expressed in non-infected bystander cells. We did not observe an inflammatory signature during infection. In conclusion, iPSC-HLCs are an immune competent platform to study responses to EBOV infection.

Neovasculogenic effect of 11,12-epoxyeicosatrienoic acid involves the Akt/eNOS signaling pathways in human endothelial progenitor cells.

The 11,12-epoxy-eicosatrienoic acid (11,12-EET) is formed from arachidonic acid (AA) by cytochrome P450 2J2 (CYP 2J2) epoxygenase and function as an effector in blood vessels. Human endothelial progenitor cells (hEPCs), a preceding cell source for endothelial cells (ECs), involve in the vascular tissue repairing by postnatal neovasculogenesis. However, the effect of 11, 12-EET on hEPCs and neovasculogenesis is not well known. In the current study, we examined the function of 11, 12-EET in hEPCs-mediated neovasculogenesis by using tubular formation analysis, Western Blotting assay, immunofluorescence staining, flow cytometry analysis and zymogram analysis. The results suggest that 11, 12-EET significantly induces neovasculogenesis through the phosphorylation of phosphoinositide 3-kinase (PI3-K)/Akt, endothelial-nitric oxide synthase (e-NOS) and extracellular signal-regulated kinase 1/2 (ERK 1/2) signaling pathways. 11, 12-EET up-regulates the expression of cyclin D1, cyclin-dependent kinase 4 (CDK4) and nuclear factor kappa B (NF-κB) proteins. Moreover, 11, 12-EET augments the expression of VE-cadherin and CD31 proteins in hEPCs. 11, 12-EET also augmented Rac1/Rho A signaling cascades, cell migration and an up-regulation of matrix metalloproteinase (MMP) -2 and -9 proteins. These results demonstrate that 11, 12-EET exerts a significant function in the neovasculogenesis of hEPCs.

Label-free chemical imaging of cytochrome P450 activity by Raman microscopy

Although investigating drug modulation of cytochrome P450 (CYP) activity under physiological conditions is crucial in drug development to avoid severe adverse drug reactions, the current evaluation approaches that rely on the destructive and end-point analysis can be misleading due to invasive treatments and cellular heterogeneity. Here, we propose a non-destructive and high-content method for visualizing and quantifying intracellular CYP activity under drug administration by Raman microscopy. The redox-state and spin-state sensitive Raman measurement indicated that the induced CYPs in living hepatocytes were in oxidized and low-spin state, which is related to monooxygenase function of CYP. Moreover, glycogen depletion associated with CYP induction was simultaneously observed, indicating a relevant effect on glucose metabolism. By deciphering the overall changes in the biochemical fingerprints of hepatocytes, Raman microscopy offers a non-destructive and quantitative chemical imaging method to evaluate CYP activity at the single-cell level with the potential to facilitate future drug development schemes.

Knockdown of CYP6CR2 and CYP6DE5 reduces tolerance to host plant allelochemicals in the Chinese white pine beetle Dendroctonus armandi

Bark beetles rely on detoxifying enzymes to resist the defensive terpenoids of the host tree. Insect cytochrome P450 (CYPs) plays a key role in the detoxification of pesticides and plant allelochemicals. CYP6 family is unique to Insecta, and its biochemical function is basically related to the metabolism of exogenous substances. In this study, we sequenced and characterized the full-length cDNAs of two CYP6 genes from Chinese white pine beetle, Dendroctonus armandi. Spatiotemporal expression profiling revealed that the expression of CYP6CR2 and CYP6DE5 was higher in larval and adult stages of D. armandi than that in other developmental stages, and that two genes predominantly expressed in brain, midgut, fat body, Malpighian tubules or hemolymph. The expression of CYP6CR2 and CYP6DE5 was significantly induced after feeding on the phloem of Pinus armandii and exposure to six stimuli [(±)- α -pinene, (−)-α-pinene, (−)-β-pinene, (+)-3-carene, (±)-limonene and turpentine]. Importantly, silencing CYP6CR2 and CYP6DE5 separately could increase the sensitivity, led to a significant reduction of the activity of P450, resulting a significant increase in adult mortality after treatment with terpenoids. The comprehensive results of this study showed that in the process of host selection and colonization, the functions of CYPs were mainly to hydrolyze the chemical defense of the host and degrade odor molecules. These findings may help to develop new treatments to control this important pest.

Human Cytochrome P450 8B1 Structure and Function: Supporting Drug Design for NAFLD and Diabetes Mellitus

Human cytochrome P450 8B1 (CYP8B1) performs a key step in human production of bile acids. It 12α‐hydroxylates a steroidal substrate to produce the relatively polar primary bile acid cholic acid. The absence of CYP8B1 catalytic function results in the more hydrophobic bile acid chenodeoxycholic acid. Knockouts of CYP8B1 indicated this P450 may be a good drug target for nonalcoholic fatty liver disease due to its alteration of bile acid‐mediated fat absorption. Similar studies indicate that the absence of CYP8B1 also has a favorable effect on type 2 diabetes. However, the absence of selective CYP8B1 inhibitors both restricts further study of its physiological roles and potential disease treatment. We recently generated the first CYP8B1 structure, with the nonspecific type II inhibitor (S)‐tioconazole, and are now employing a structure/function strategy to understand CYP8B1 interactions with a) its native substrate and b) a wider array of inhibitors. Within the CYP8B1 active site tioconazole is located near the B’ helix due to spatial hinderance from W281 and N286 in the I helix on the opposite side of the active site. W281 has previously been suggested as a key residue for CYP8B1 catalysis with its native steroidal substrate. To test this concept, a human CYP8B1 W281F mutant was generated. The W281F enzyme demonstrated both impaired substrate binding and significantly reduced catalytic efficiency. Thus W281 appears to be important in binding both the native substrate and the tioconazole inhibitor. Subsequently a set of additional azole ligands were evaluated to broaden the definition of ligand structures compatible with the CYP8B1 active site and potentially identify even better inhibitors. Changes in visible absorbance spectra were used to determine the binding mode and determine Kd values, while inhibition of steroid 12a‐hydroxylation was used to determine IC50 values. Most azole compounds bound CYP8B1, typically via the expected type II imidazole‐to‐heme‐iron interaction like tioconazole, and with Kd values in the nM to µM range. Miconazole and econazole bound with the highest affinity and had the lowest IC50 values, demonstrating improvements over tioconazole. The structural differences between these ligands are a relatively moderate substitution of a thienyl vs. phenyl ring and modified chlorination patterns on that ring, demonstrating CYP8B1 sensitivity to these inhibitor components. Both the CYP8B1 structure and the functional characterization should facilitate the identification increasingly effective CYP8B1 inhibitors. If selectivity can also be incorporated into these compounds, the resulting inhibitors should enable a better understanding of the role of CYP8B1 in normal physiology and facilitate a potential treatment for nonalcoholic fatty liver disease and type 2 diabetes.

Identification and analysis of CYP450 family members in Ginkgo biloba reveals the candidate genes for terpene trilactone biosynthesis

Cytochrome P450 (CYP450), a superfamily of mono-oxygenases, participates in various physiological processes of plants. Terpene trilactones (TTLs) are unique bioactive components of Ginkgo biloba L. and have been applied in many fields including industry, medicine, and health. In this study, a total of 187 CYP450s were identified from the ginkgo genome and were divided into six clans, of which the CYP71 clan (10 subfamilies) included all A-type CYP450s, with a total of 83. The other five clans (18 subfamilies) contained 104 non-A-type CYP450s. The physicochemical properties, functional domains, conserved motifs, phylogenic relationships, gene structures, and chromosome locations of the GbCYP450s were comprehensively analyzed. The GbCYP450s had a conserved domain of FXXGXRXCXG, and each clan or subfamily had its own characteristics, indicating potential differences in their functions. A total of 114 identified differentially expressed CYP450s were identified from transcriptome sequencing. Combined with the content of TTLs, 22 CYP450s that might be involved in TTL biosynthesis were screened via correlation and weighted gene correlation network analysis. By constructing a phylogenetic tree with CYP450s involved in terpenoid biosynthesis in other plants, and 22 CYP450s had a close evolutionary relationship, which further verified our hypothesis. Additionally, the MeJA treatment could increase the content of TTLs in G. biloba, and 12 of the 22 candidate GbCYP450s contained cis-acting regulatory elements involved in methyl jasmonate (MeJA)-responsiveness, among which eight CYP450s genes were strongly induced by MeJA to up-regulate expression. This study provides a foundation for clarifying the function of CYP450s in TTL biosynthesis and the use of genetic engineering technology to increase the production of TTLs.

Genome-wide identification, evolution analysis of cytochrome P450 monooxygenase multigene family and their expression patterns during the early somatic embryogenesis in Dimocarpus longan Lour.

Cytochrome P450 (CYP), a multi-gene superfamily, is involved in a broad range of physiological processes, including hormone responses and secondary metabolism throughout the plant life cycle. Longan (Dimocarpus longan), a subtropical and tropical evergreen fruit tree, its embryonic development is closely related to the yield and quality of fruits. And a large number of secondary metabolites, such as flavonoids and carotenoids, are also produced during the longan somatic embryogenesis (SE). It is important, therefore, to study potential functions of CYPs in longan. However, the knowledge of longan CYPs is still very limited. Here, a total of 327 DlCYPs were identified using the genome-search method, which could be classified into nine clans. The expansion of the DlCYP family was mainly caused by tandem duplication (TD) events. Promoter cis-acting elements analysis elucidated that DlCYPs played important roles in hormonal responses. A total of 246 DlCYPs exhibited six different expression patterns during the early SE based on longan transcriptomic data. Eight DlCYPs underwent alternative splicing (AS) events, and they might produce one to six isoforms. And the AS transcript of DlCYP97C1 might act as an alternative to the full-length transcript in ICpEC and GE stages. Finally, protein–protein interaction (PPI) networks and miRNA target prediction elucidated that DlCYPs might be involved in the phenylpropanoid metabolic pathway and primarily regulated and targeted by miR413. In summary, our results provided valuable inventory for understanding the classification and biological functions of DlCYPs and provided insight into further functional verification of DlCYPs during the longan early SE.

The Role of Cytochrome P450 Enzymes in COVID-19 Pathogenesis and Therapy.

Coronavirus disease 2019 (COVID-19) has become a new public health crisis threatening the world. Dysregulated immune responses are the most striking pathophysiological features of patients with severe COVID-19, which can result in multiple-organ failure and death. The cytochrome P450 (CYP) system is the most important drug metabolizing enzyme family, which plays a significant role in the metabolism of endogenous or exogenous substances. Endogenous CYPs participate in the biosynthesis or catabolism of endogenous substances, including steroids, vitamins, eicosanoids, and fatty acids, whilst xenobiotic CYPs are associated with the metabolism of environmental toxins, drugs, and carcinogens. CYP expression and activity are greatly affected by immune response. However, changes in CYP expression and/or function in COVID-19 and their impact on COVID-19 pathophysiology and the metabolism of therapeutic agents in COVID-19, remain unclear. In this analysis, we review current evidence predominantly in the following areas: firstly, the possible changes in CYP expression and/or function in COVID-19; secondly, the effects of CYPs on the metabolism of arachidonic acid, vitamins, and steroid hormones in COVID-19; and thirdly, the effects of CYPs on the metabolism of therapeutic COVID-19 drugs.

Analysis of CYP450 gene expression and function in white-rot fungus, Lenzites gibbosa, treated with Congo red

ABSTRACT We investigated the function of cytochrome P450 (CYP450) genes in degradation of the diazo dye, Congo red, by white-rot fungus Lenzites gibbosa. Hyphae treated with Congo red at different times were sequenced to obtain transcription data. CYP450 genes in transcriptomes were identified using a gene-encoding protein functional search and analyzed in Cluster of Orthologous Genes, Gene Ontology, Kyoto Encyclopedia of Genes and Genomes Databases. Differentially expressed genes (DEGs) in different groups were analyzed using EdgeR. We present the relation between transcription factors (TFs) and CYP450 regulation by analysis of the co-expression network. One hundred sixty CYP450 genes obtained by a functional annotation search were related to the oxido-reduction reaction of secondary metabolism, defense mechanism and aromatic compound degradation. The fastest decolorization and the greatest expression of CYP450 genes, which were related to the decolorization effect, occurred at 0–3 h. Seven CYP450 genes (7522, 6568, 4482, 9118, 10935, 7521 and 10926) were identified. The key TFs that regulate these genes belong to the zinc finger family. CYP450 genes and their products in L. gibbosa participated in degradation of Congo red and stress resistance. We provide a reference value for degradation and decolorization of Congo red.

Fish health in the Nidda as an indicator for ecosystem integrity: a case study for Central European small streams in densely populated areas

BackgroundIn Germany and the EU, most headwaters are still far from reaching a good chemical and ecological status as it is required by the European Water Framework Directive (WFD), until 2027 the latest. Particularly, in densely populated areas, impacts from municipal and industrial wastewater discharges or diffuse agricultural emissions are still a matter of concern. This also applies to the Nidda River which is considered to be in a moderate to rather poor condition. In our study, we investigated short-term and long-term consequences of anthropogenic pollution on fish health via one monitoring with caged fish (CF) and two field sampling campaigns (FF). In the CF monitoring, rainbow trout (Oncorhynchus mykiss) were caged for seven weeks at four selected sites along the Nidda, whereas in the FF monitoring approach, feral fish, including brown trout (Salmo trutta f. fario), European chub (Leuciscus cephalus) and stone loach (Barbatula barbatula) were caught in June and September 2016.ResultsHistopathological analyses of liver and gills were conducted, accompanied by measurements of hepatic 7-ethoxyresorufin-O-deethylase (EROD) activity to assess the cytochrome P450 (CYP1A1) function, and genotoxicity via the micronucleus assay. Caged as well as field-captured fish exhibited impaired health conditions showing lesions particularly in the liver, and a presumably overwhelmed CYP1A1 system, whereas genotoxicity was not induced. The variation between sampling sites and seasons was rather low, but two trends were recognisable: (a) liver condition was poorest around spawning season and (b) tissue integrity and EROD activity were most affected downstream of industrial dischargers. Furthermore, effects were species dependent: the generally highly sensitive S. trutta f. fario proved to be impacted most, whereas L. cephalus with its pelagic lifestyle was affected less than the benthic B. barbatula, indicating a relevant contamination of sediments.ConclusionOur results confirm the impaired ecological state of the Nidda and emphasise that a sustainable improvement of aquatic ecosystem health needs to include both water quality and sediment contamination to approach the ambitious WFD goal.